Corneal epithelial cells migrate and proliferate;importantly they do so directionally, to heal wounds. Growth factors and cytokines play pivotal roles in wound healing and may be potential targets for corneal wound therapies. We have discovered a very different factor, namely naturally-occurring electric fields (EFs) at corneal wounds that also activate intracellular pathways. More significantly, because EFs are intrinsically directional, they activate signaling pathways directionally, giving cells a directional cue and guiding cells to migrate and divide in the direction of the wound to facilitate healing. Our studies have shown that EFs override other well-accepted directional cues such as contact inhibition release, wound void, population pressure and chemotaxis to guide cell migration in a defined direction. How the endogenous EFs are generated and regulated is not known. In streptozotocin (STZ)- induced type 1 diabetes mellitus rats and Pax6+/- mutant mice that have defective corneal wound healing, we observed significantly reduced endogenous wound EFs. Can we enhance the endogenous wound EFs to enhance wound healing, especially in refractory and chronic wounds? Our long-term goal is to elucidate the mechanisms through which electric signals can be exploited to accelerate wound healing. We recently observed that wound EFs increase gradually following injury, and substitution of Cl- or Na+ in the bathing solution significantly alters the endogenous EFs. We thus hypothesize that injury to the cornea induces actively-regulated wound electric fields, which are formed by fluxes of specific ions (e.g. Cl-) that are controlled by Cl- channels and transport molecules;manipulating Cl- flux may enhance endogenous electric fields and wound healing. We will test this hypothesis with the following Specific Aims: Aim 1. To confirm that wound electric fields are an active response to injury. Aim 2. To determine the ionic mechanisms of endogenous EFs at corneal wounds. Aim 3. To elucidate the molecular mechanisms of wound electric fields. The results from this application will define the active electric signaling in corneal wound healing, provide ionic and molecular mechanisms of electric signaling in wound healing, and may lead to novel therapies to improve wound healing exploiting electric signaling. PUBLIC HEALTH RELEVANCE: Persistent corneal epithelial defects pose an important medical problem. We recently discovered a novel signaling mechanism at corneal wounds, namely naturally-occurring electric fields that have profound guidance effects on epithelial cells to heal wounds. This project seeks to determine the ionic and molecular mechanisms controlling ionic fluxes at corneal wounds. The scientific knowledge to be acquired through this project, i.e. how cells regulate this fundamental signal, will open a new avenue to treat delayed and non-healing corneal wounds, and wounds in general.